• Title/Summary/Keyword: protein protein interactions

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Ig G fusion 단백질을 사용한 리간드-수용체의 상호작용

  • 천혜경
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.11a
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    • pp.143-145
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    • 1994
  • Chimeric fusion proteins involving IgG have proven valuable in studying protein-protein interactions and may possess therapeutic applications as well. For example, three receptor subtypes for the natriuretic peptides, when fused to the Fc portion of human IgG ${\gamma}$ chain, were quantitatively and qualitatively indistinguishable from the native receptor, thus allowing detailed structure-function studies of the receptor. In an attempt to block human immunodeficiency virus infectivity with soluble derivatives of CD4, a CD4/IgG Fc chimeric molecule was shown to increase the plasma half life of soluble CD4 and possessed the added advantage of IgG Fc-mediated placental transfer. In the case of the KGFR, this approach provided a framework for dissection of its ligand binding domains and made it possible to demonstrate that high affinity binding sites for two ligands, aFGF and KGF, reside within different receptor Ig-like domains. Chimeric molecules fused to immunoglobulins would have the advantages of secretion from transfected cells as well as detection and purification from medium utilizing Staphylococcus aureus Protein A. In addition, where highly related receptors make their discrimination very hard due to the difficulties in generating specific immunochemical probes, IgG fusion protein with tailor-made specificities confers particular advantages to elucidate patterns of receptor distribution and expression. The approach described here may have general applications in defining ligand-receptor interactions as well as searching for specific agonists and antagonists of receptor function.

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P22-Based Challenge Phage Constructs to Study Protein-Protein Interactions between the $\sigma$$^{54}$-Dependent Promoter, dctA, and Its Transcriptional Regulators

  • Song, Jeong-Min;Kim, Eungbin;Lee, Joon H.
    • Journal of Microbiology
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    • v.40 no.3
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    • pp.205-210
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    • 2002
  • To study interactions between $C_{4}$-dicarboxylic acid transport protein D and E$\sigma$$^{54}$ in the dctA promoter regulatory region, we used the challenge phage system. An ant'-`lac fusion was recombined onto the challenge phage, and this ant'-`lac fusion along with Pant and the R. meliloti dctA promoter regulatory region were cloned onto a plasmid. The plasmid bearing the ant'-`lac fusion was used as a reporter plasmid in a coupled transcription-translation system. Addition of purified $\sigma$$^{54}$ to the coupled system specifically repressed transcription of the plasmid-borne ant'-`lac fusion. When DCTD was added along with $\sigma$$^{54}$ to the coupled system, transcription of the ant'-`lac fusion was even further repressed, suggesting that DCTD may stabilize closed complexes between E$\sigma$$^{54}$ and the dctA promoter.

A Cell-based Method to Monitor the Interaction between Hepatitis B Virus Capsid and Surface Proteins

  • Kim, Yun-Kyoung;Oh, Soo-Jin;Jin, Bong-Suk;Park, Chan-Hoo;Jeon, Hye Sung;Boo, Doo-Wan;Yu, Yeon-Gyu
    • Bulletin of the Korean Chemical Society
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    • v.30 no.3
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    • pp.577-581
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    • 2009
  • Interactions between the surface and capsid proteins of the hepatitis B virus (HBV) are critical for the assembly of virus particles. In this study, we developed a cell-based method to visualize the interactions between the capsid and surface proteins of HBV. Capsid-GFP, a capsid protein fused to a green fluorescence protein (GFP), forms nucleocapsid-like structures in the cytoplasm of mammalian cells. It relocates to the plasma membranes in cells expressing PH-PreS, a fusion protein consisting of the PreS region of the HBV surface protein and the PH domain of PLC-$\gamma$. Membrane localization of the capsid-GFP in these cells is prevented by an inhibitory peptide that blocks the interaction between the capsid and surface proteins. This dynamic localization of capsid-GFP is applicable for screening compounds that may potentially inhibit or prevent the assembly process of HBV particles.

Effects of Morphine and Panax ginseng on the Opioid Receptor-G protein Interactions

  • Kim, Young-Ran;Kim, Ae-young;Kim, Kyeong-Man
    • Biomolecules & Therapeutics
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    • v.7 no.1
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    • pp.1-6
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    • 1999
  • Effects of Panax ginseng on the morphine toxicity were studied in relation to its effects on the opioid receptor-G protein interactions. Morphine treatments (3 days) reduced the body weight increment rate and the weight of the thymus and spleen. These changes were usually recovered by the concomitant administration of ginseng total saponin (GTS) but occasionally further deteriorated. This discrepancy was studied in relation to the opioid receptor coupling to G protein, that is, the effects of morphine and GTS on the opioid receptors were studied using the antagonist-agonist competitive binding studies. When GTS recovered the morphine toxicity, morphine shifted the striatal $\delta$ receptors to slightly higher affinity state, and this was partly recovered by the GTS treatment. However, morphine did not have any effect on the affinity state of $\delta$ receptor from NG108-15 cells, suggesting that additional factors were needed for the modulation of the affinity states of $\delta$ receptor. Effects of morphine and GTS on $\mu$ receptor were complicate and variable, and we could not reach a clear conclusion. The morphine toxicity might accompany complicate biological involvements, and the modulation of the affinity states of the opioid receptors might explain a part of the effects of GTS on the morphine toxicity.

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The Changes of Phytic Acid Content and its Interactions with Protein and Minerals in the Preparation of Tempeh (Tempeh 제조시 Phytic Acid 함량변화 및 그에 따른 단백질, 무기질과의 상호작용에 관한 연구)

  • Park, Eun-Soon;Yoon, Sun
    • Journal of Nutrition and Health
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    • v.16 no.4
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    • pp.281-286
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    • 1983
  • The interactions of phytic acid with protein and minerals has been blamed to decrease the bioavailability of protein and minerals in soybean products. Tempeh, the traditional Infonesian fermented soybean product, was prepared to investigate the changes of phytic acid contents nesian fermented soybean product, was prepared to investigate the changes of phytic acid contents and its interactions with protein and minerals in the fermentation. The acceptability of tempeh were also studied by conducting sersory evaluation. 1) Phytic acid contents of cooked soybeans and of tempch were significantly lower than that of raw soybeans, indicating that cooking and fermentation resulted in the decrease in phytic acid content of soybeans. In tempeh the fraction of phytic acid retained after ultrafiltration was significantly lower than that in raw soybeans. 2) The total protein contents were not significantly different between raw soybeans and tempeh. Phytic acid contents per gram of protein retained ultrafiltration were significantly higher in raw soybeans than in tempeh. This result is interpreted as that raw soybeans contain higher amounts of phytic acid- protein complexes than tempeh. 3) Both of calcium and zinc contents were not significantly different among raw, cooked soybeans and tempeh. However, the retained Ca and Zn fraction after ultrafiltration were significantly lower in tempeh comparing with that in raw soybeans. Lower retention of Ca and Zn after ultrafiltration in tempeh may be the result of lower phytate content of tempeh, thereby less chance of forming mineral- phytate complexes. 4) Tempeh received the sensory evaluation scores between good and fair and the addition of garlic to tempeh significantly improved the odor, general desirability and total score.

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Surface Interactions of Model Peptides for Mussel Adhesive Protein

  • Cho, Eun-Kyung;Lee, Yoon-Hee;Cho, Nam-Jun;Cha, Hyung-Joon
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.630-633
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    • 2003
  • The mussel adhesive protein Mefp-1 is a natural, strong and durable adhesive that is stable under corrosive, saline conditions. Mefp-1 is found in the marine mussel Mytilus edulis and it has a molecular weight of ca. 130,000. The primary structure is mainly composed of repeating decapetides: Ala-Lys-Pro -Ser-Tyr Hyp-Hyp-Thr-DOPA-Lys. To elucidate the mechanism by which Mefp-1 bonds to metal surfaces, we have used surface-enhanced Raman spectroscopy to study the interactions of peptides related to the Mefp-1 decapeptide repeat with gold surfaces. We have concluded that the tyrosine residue and the carboxyl terminus interact strongly with the gold surface, and that proline and hydroxyproline constrain the conformations of the peptides, thereby limiting the types of possible interactions of the functional groups with the gold surface.

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Involvement of Electrostatic Interactions between the Components of Toluene Dioxygenase from Pseudomonas putida Fl

  • Lee, Kyoung
    • Journal of Microbiology and Biotechnology
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    • v.8 no.4
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    • pp.416-421
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    • 1998
  • A carboxyl group modifier, l-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) was used to study the interactions between three components of toluene dioxygenase (TDO) from Pseudomonas putida FI. $Ferredoxin_{TOL}$ activity was increased by the treatment with EDC; however, the activity was rapidly declined in the prolonged incubation. In covalent cross-linking experiments with EDC, $Ferredoxin_{TOL}$ made a one-to-one complex with $Reductase_{TOL}$ or the large subunit of $ISP_{TOL}$. These results provide evidence for the involvement of electrostatic interactions in the TDO electron transfer system.

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Extraction of Protein-Protein Interactions based on Convolutional Neural Network (CNN) (Convolutional Neural Network (CNN) 기반의 단백질 간 상호 작용 추출)

  • Choi, Sung-Pil
    • KIISE Transactions on Computing Practices
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    • v.23 no.3
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    • pp.194-198
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    • 2017
  • In this paper, we propose a revised Deep Convolutional Neural Network (DCNN) model to extract Protein-Protein Interaction (PPIs) from the scientific literature. The proposed method has the merit of improving performance by applying various global features in addition to the simple lexical features used in conventional relation extraction approaches. In the experiments using AIMed, which is the most famous collection used for PPI extraction, the proposed model shows state-of-the art scores (78.0 F-score) revealing the best performance so far in this domain. Also, the paper shows that, without conducting feature engineering using complicated language processing, convolutional neural networks with embedding can achieve superior PPIE performance.

Effects of Protein and Iron Concentrations on Iron Solubility in Black Tea Infusion (단백질 함량 및 철분 농도의 변화에 따라 홍차 추출물이 철분의 용해도에 미치는 영향)

  • 김희선
    • Journal of Nutrition and Health
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    • v.29 no.8
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    • pp.861-866
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    • 1996
  • Tannins in plant foods and beverages may produce antinutritional or toxic effects although some proteins with high affinity for tannins seem to function as defense mechanism to tannin toxicity. Our objectives were to investigate of tea tannins, iron and protein and to evaluate the role of proteins in tannin effects on iron solubility. Iron solubility in vitro was measured using tea with and without proteins. Mixtures of tea, protein in varying concentrations(either gelatin or bovine serum albumin), and iron(eithe 10 or 50ug/mL) were prepared. Controls contained water in place of tea. Iron bioavailability was assessed by measuring iron solubility in the simulated gastric condition with pepsin digestion. Bound iron was removed by centrifugation and soluble in tea alone. When iron concentratin was 10ug/mL, addition of small amounts of protein to tea dramatically reduced iron solubility, but solubility of iron increased in the tea mixturea as the concentration of protein was increased. The percnetage of iron that precipitated was much greater at 10ug Fe/mL than the values at 50ug Fe/mL suggesting that the iron binding sites on the tea-protein complex was saturated. These results suggest that interactions of iron with tea tannins are influenced by the concentratins of protein and iron.

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Construction and Characterization of an Enhanced GFP-Tagged TIM-1 Fusion Protein

  • Qing, Jilin;Xiao, Haibing;Zhao, Lin;Qin, Guifang;Hu, Lihua;Chen, Zhizhong
    • Journal of Microbiology and Biotechnology
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    • v.24 no.4
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    • pp.568-576
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    • 2014
  • TIM-1 (also known as KIM-1 and HAVcr-1) is a type I transmembrane glycoprotein member of the TIM family that may play important roles in innate and adaptive immune responses. The overexpression of proteins associated with membrane proteins is a major obstacle to overcome in studies of membrane protein structures and functions. In this study, we successfully coupled the overexpression of the TIM-1 protein with a C-terminal enhanced green fluorescent protein (GFP) tag in Escherichia coli. To the best of our knowledge, this report is the first to describe the overexpression of human TIM-1 in E. coli. The purified TIM-1-EGFP fusion protein recognized and bound directly to apoptotic cells and did not to bind to viable cells. Furthermore, we confirmed that the interactions of TIM-1-EGFP with apoptotic cells were blocked by TIM-1-Fc fusion proteins. This fusion protein represents a readily obtainable source of biologically active TIM-1 that may prove useful in future studies of human TIM-1.